Ribonucleases (RNases), essential for RNA metabolism, are implicated in human diseases, including neurodevelopmental, developmental, hematopoietic and other dysfunctions through mutations that disrupt their enzymatic ...Ribonucleases (RNases), essential for RNA metabolism, are implicated in human diseases, including neurodevelopmental, developmental, hematopoietic and other dysfunctions through mutations that disrupt their enzymatic functions. Exploring RNase mutations across organisms offers insights into Mendelian diseases, facilitating molecular dissection of pathological pathways and therapeutic development. By employing model organisms, our analysis underscores the evolutionary conservation of RNase genes, facilitating deeper insights into disease mechanisms. These models are vital for uncovering rare molecular dysfunctions and potential therapeutic targets, demonstrating the effectiveness of integrated research approaches in addressing complex genetic disorders. Drawing from phylogenetic analyses, literature survey, and databases documenting the effects of human disease-causing mutations, the review highlights the significance and advantages of employing model organisms to study specific Mendelian disorders.展开更多
Self-incompatibility is an intraspecific reproductive barrier to prevent self-fertilization inthe flowering plants. In many species, self-incompatibility is controlled by a single S locus with multiple alleles. So far...Self-incompatibility is an intraspecific reproductive barrier to prevent self-fertilization inthe flowering plants. In many species, self-incompatibility is controlled by a single S locus with multiple alleles. So far, the only gene known in the S locus of the Solanaceae, Scrophulariaceae and Rosaceae encodes a class of ribonucleases, called self-incompatibility ribonucleases (S RNases), which have been shown to mediate stylar expression of self-incompatible reaction. As the first step to investigate their three-dimensional structure, we successfully expressed three biologically active S RNases of Antirrihnum (S2, S4 and S5) in Escherichia coli (E. coli). Their functional expressions caused no detrimental effect on host bacteria growth and provided a basis for a large scale preparation of S RNase proteins. Possible reasons for non-lethality of S RNases on E. coliare discussed.展开更多
To explore the functions of human ribonuclease 9(RNase 9),we constructed a mammalian fusion expression vector pcDNA-hRNase9,prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N...To explore the functions of human ribonuclease 9(RNase 9),we constructed a mammalian fusion expression vector pcDNA-hRNase9,prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N-terminal amino acid sequences.According to the determined mature protein,recombinant human RNase 9 was prepared in E.coli.Ribonucleolytic activity and antibacterial activity of recombinant human RNase 9 were detected,and the distribution of human RNase 9 on tissues and ejaculated spermatozoa and in vitro capacitated spermatozoa were analyzed via indirect immunofluorescence assay.The results showed that recombinant human RNase 9 did not exhibit detectable ribonucleolytic activity against yeast tRNA,but exhibited antibacterial activity,in a concentration/time dependent manner,against E.coli.Immunofluorescent analyses showed that the predicted human RNase 9 was present throughout the epididymis,but not present in other tissues examined,and human RNase 9 was also present on the entire head and neck regions of human ejaculated spermatozoa and in vitro capacitated spermatozoa.These results suggest that human RNase 9 may play roles in host defense of male reproductive tract.展开更多
Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode...Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode immunoassay was developed to detect OPs(triazophos,parathion,and chlorpyrifos)in apples,turnips,cabbages,and rice.Gold nanoparticles were functionalized with monoclonal antibodies against the tested OPs.DNA oligonucleotides were complementarily hybridized with an RNA fluorescent label for signal amplification.The detection signals were generated by DNA-RNA hybridization and ribonuclease H dissociation of the fluorophore.The resulting fluorescence signal enables multiplexed quantification of triazophos,parathion,and chlorpyrifos residues over the concentration range of 0.01-25,0.01-50,and 0.1-50 ng/mL with limits of detection of 0.014,0.011,and 0.126 ng/mL,respectively.The mean recovery ranged between 80.3% and 110.8% with relative standard deviations of 7.3%-17.6%,which correlate well with results obtained by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The proposed bio-barcode immunoassay is stable,reproducible and reliable,and is able to detect low residual levels of multi-residue OPs in agricultural products.展开更多
Members of the Pathogenesis Related(PR)10 protein family have been identified in a variety of plant species and a wide range of functions ranging from defense to growth and development has been attributed to them.PR10...Members of the Pathogenesis Related(PR)10 protein family have been identified in a variety of plant species and a wide range of functions ranging from defense to growth and development has been attributed to them.PR10 protein possesses ribonuclease(RNase)activity,interacts with phytohormones,involved in hormone-mediated signalling,afforded protection against various phytopathogenic fungi,bacteria,and viruses particularly in response to biotic and abiotic stresses.The resistance mechanism of PR10 protein may include activation of defense signalling pathways through possible interacting proteins involved in mediating responses to pathogens,degradation of RNA of the invading pathogens.Moreover,several morphological changes have been shown to accompany the enhanced abiotic stress tolerance.In this review,the possible mechanism of action of PR10 protein against biotic and abiotic stress has been discussed.Furthermore,our findings also confirmed that the in vivo Nitric oxide(NO)is essential for most of environmental abiotic stresses and disease resistance against pathogen infection.The proper level of NO may be necessary and beneficial,not only in plant response to the environmental abiotic stress,but also to biotic stress.The updated information on this interesting group of proteins will be useful in future research to develop multiple stress tolerance in plants.展开更多
Objective：The association between ribonuclease L（RNASEL）gene polymorphisms and prostate cancer risk has been widely reported,but the results of these studies remained controversial and underpowered.We performed a m...Objective：The association between ribonuclease L（RNASEL）gene polymorphisms and prostate cancer risk has been widely reported,but the results of these studies remained controversial and underpowered.We performed a meta-analysis of 28 studies to evaluate the association between Arg462Gln and Asp541Glu polymorphisms in the RNASEL gene and prostate cancer risk.Methods：Odds ratios（ORs）with 95%confidence intervals（CIs） were estimated to assess the association between RNASEL polymorphisms and prostate cancer risk.Results：A significantly increased prostate cancer risk was found for the Arg462Gln polymorphism in Africans（Gln/Gln vs Arg/Arg：OR=2.50,95%CI=1.28-4.87;Gln/Gln vs Gln/Arg＋Arg/Arg：OR=2.54,95%CI=1.30-4.95）,but not in Europeans and Asians.Additionally,the Asp541Glu polymorphism was associated with increased total prostate cancer risk（Glu-allele vs Asp-allele：OR=1.04,95%CI=1.01-1.07;Glu/Glu vs Asp/Asp：OR=1.22,95%CI= 1.03-1.46;Glu/Glu vs Glu/Asp＋Asp/Asp：OR=1.09,95%CI=1.02-1.16）.In the stratified analysis for the Asp541Glu polymorphism,there was a significantly increased prostate cancer risk in Africans and Europeans,and in hospital-based prostate cancer cases.Conclusion：The meta-analysis results showed evidence that RNASEL Arg462Gln and Asp541Glu polymorphisms are associated with prostate cancer risk and could be low-penetrance prostate cancer susceptibility biomarkers.展开更多
Male infertility is a major reproductive disorder,which is clinically characterized by highly heterogeneous phenotypes of abnormal sperm count or quality.To date,five male patients with biallelic loss-of-function(LOF)...Male infertility is a major reproductive disorder,which is clinically characterized by highly heterogeneous phenotypes of abnormal sperm count or quality.To date,five male patients with biallelic loss-of-function(LOF)variants of PARN-like ribonuclease domain-containing exonuclease 1(PNLDC1)have been reported to experience infertility with nonobstructive azoospermia.The aim of this study was to identify the genetic cause of male infertility with oligo-astheno-teratozoospermia(OAT)in a patient from a Chinese Han family.Whole-exome and Sanger sequencing analyses identified a homozygous LOF variant(NM_173516.2,c.l42C>T,p.Gln48Ter)in PNLDC1.Hematoxylin and eosin staining revealed that the spermatozoa of the patient with OAT had an irregular head phenotype,including microcephaly,head tapering,and globozoospermia.Consistently,peanut agglutinin staining of the spermatozoa revealed a complete or partial loss of the acrosome.Furthermore,the disomy rate of chromosomes in the patient’s spermatozoa was significantly increased compared with that of a fertile control sample.We reported an LOF variant of the PNLDC1 gene responsible for OAT.展开更多
BACKGROUND Bioinformatics analysis showed that the expression of the poly(A)-specific ribonuclease(PARN)gene in gastric cancer,head and neck squamous cell carcinoma,melanoma,cervical cancer and lung squamous cell carc...BACKGROUND Bioinformatics analysis showed that the expression of the poly(A)-specific ribonuclease(PARN)gene in gastric cancer,head and neck squamous cell carcinoma,melanoma,cervical cancer and lung squamous cell carcinoma tissues was significantly higher than that in normal tissues and was associated with high stage and poor prognosis.The expression of the PARN gene in esophageal cancer(EC)tissue is also significantly higher than that in normal tissues,but the effect of PARN on the proliferation,migration and invasion of EC cells remains unclear.AIM To investigate the relationship between PARN and the proliferation,migration and invasion of EC cells.METHODS The EC tissues of 91 patients after EC surgery and 63 paired precancerous healthy tissues were collected.PARN mRNA levels were measured using a tissue microarray,and the PARN expression level was evaluated using immunohistochemistry to analyze the relationship between PARN expression and clinicopathologic features as well as the survival and prognosis of patients.In addition,the effects of PARN gene knockout on tumor cell proliferation,invasion and migration were studied by using shRNA during the in vitro culture of EC cell lines Eca-109 and TE-1,and the effects of the PARN gene on tumor growth in vivo were verified by a xenotransplantation nude mice model.RESULTS The expression of PARN in EC tissues was higher than that in adjacent normal tissues,and the level of PARN expression was significantly positively correlated with lymphatic metastasis.Patients with high PARN levels had poor overall survival.BIM,IGFBP-5 and p21 levels were significantly increased in the PARN knockout group,while the expression levels of the antiapoptotic proteins Survivin and sTNF-R1 were significantly decreased in the apoptotic antibody array data.In addition,the expression levels of Akt,p-Akt,PIK3CA and CCND1 in the downstream signaling pathway regulating EC progression were significantly decreased.The culture of EC cell lines confirmed that the apoptosis rate of EC cells was significantly increased,the growth and proliferation of tumor cells were significantly inhibited,and the invasion and migration ability of tumor cells were significantly decreased after PARN gene knockout.In vivo experiments of BALB/c nude mice transfected with Eca-109 cells expressing control shRNA(sh-NC)and PARN shRNA(sh-PARN)showed that the tumor volume and weight of nude mice treated with sh-PARN were significantly decreased compared with those of nude mice treated with sh-NC,indicating that PARN knockdown significantly inhibited tumor growth in vivo.CONCLUSION PARN has antiapoptotic effects on EC cells and promotes their proliferation,invasion and migration,which is associated with the development of EC and poor patient prognosis.PARN may become a potential target for the diagnosis,prognosis prediction and treatment of EC.展开更多
Early stage expression of PR10 combined with phytoalexins contributed to Verticillium wilt resistance in cotton. In order to analysis the activities of PR10 proteins during pathogens’ infection, we cloned a Verticill...Early stage expression of PR10 combined with phytoalexins contributed to Verticillium wilt resistance in cotton. In order to analysis the activities of PR10 proteins during pathogens’ infection, we cloned a Verticillium-induced PR10 (GbPR10-1) gene from cotton (Gossypium barbadense) and compared its expression patterns and domains with other PR10 proteins. Bioinformatics indicated that GbPR10-1 showed the lowest similarity with other 12 different PR10 genes in cotton (Upland and sea-island cotton). Expression profiles showed that GbPR10-1 gene instantly up-regulated after infection by V. dahliae in the sea-island cotton plants. GbPR10-1 was also induced by environmental stimulus including heat, submergence and salt, and ethylene but not by ABA and salicylic acid. The GbPR10-1 protein expressed in E. coli BL21 demonstrated that it had a low ribonuclease-like activity in vitro, and could inhibit V. dahliae hyphae growth but not its spores. Comparison analysis of GbPR10-1 (from resistant species) and GhPR10-1 (from susceptible species) responding to V. dahliae infection, only GbPR10-1 gene was strongly induced in the sea-island cotton plants (incompatible response), indicating that PR10-1 genes was linked to resistance signal. In summary, the earlier activation of GbPR10-1 gene, as the index of resistance response, would be aid to block展开更多
Background: Viruses can cause different diseases in plants. To prevent viral infections, plants are treated with chemical compounds and antiviral agents. Chemical antiviral agents usually have narrow specificity, whic...Background: Viruses can cause different diseases in plants. To prevent viral infections, plants are treated with chemical compounds and antiviral agents. Chemical antiviral agents usually have narrow specificity, which limits their wide application. Alternative antiviral strategy is associated with the use of microbial enzymes, which are less toxic and are readily decomposed without accumulation of harmful substances. The aim of this work is to study the effect of Bacillus pumilus ribonuclease on various phytopathogenic viruses with specific focus on the ability of enzyme to eliminate them from plant explants in vitro. Materials and methods: Extracellular ribonuclease of B. pumilus is tested as an antiviral agent. To study the antiviral effect of RNase, depending on concentration and the time of application several plant-virus model systems are used. Virus detection is conducted by serological testing and RT-PCR. Results: Bacillus pumilus ribonuclease possesses antiviral activity against plant Rna-viruses RCMV (red clover mottle virus), PVX (Potato Virus X) and AMV (Alfalfa Mosaic Virus). The maximum inhibitory effect against actively replicating viruses is observed when plants are treated with the enzyme in the concentration of 100 ug/ml prior to infection. In case of local necrosis ribonuclease in the concentration of 1 ug/ml completely inhibits the development of RCMV virus on bean plants. The enzyme is able to penetrate plants and inhibit the development of viral infection, inhibiting effect for untreated surfaces decreased on average for 20%. It is also found that B. pumilus ribonuclease protects apical explants of sprouts of potato tubers from PVM and PVS viruses. Conclusion: B. pumilus ribonuclease possesses antiviral activity against plant Rna-viruses and produces viruses-free plants in the apical meristem culture.展开更多
2-Chlorocyclohexa-2,5-diene-1,4-dione (CBQ) or 2-chloro1,4-benzquinone is one of the common metabolites of polycyclic aromatic hydrocarbons generated through industrial processes. This report describes the biological ...2-Chlorocyclohexa-2,5-diene-1,4-dione (CBQ) or 2-chloro1,4-benzquinone is one of the common metabolites of polycyclic aromatic hydrocarbons generated through industrial processes. This report describes the biological effects of CBQ toward ribonuclease A (RNase). We also investigated the inhibition of RNase modifications and the reactivity of CBQ toward selected amino acids. The study was carried out by incubating RNase or amino acids with CBQ in a concentration- and a time-dependent manner at 37°C and pH 7.0. SDS-PAGE results showed oligomerization as well as polymeric aggregation of RNase when incubated with CBQ as early as in 10 min. CBQ-induced RNase modifications were inhibited in the presence of NADH or ascorbic acid. CBQ reactivity toward selected amino acids was also evaluated by determining the second-order rate constants for the reactions of CBQ with selected amino acids. It was found that the reactivity toward CBQ decreased in the order of lysine > threonine > serine >> aspartate > cysteine.展开更多
Herein,we studied the interaction of bovine pancreatic ribonuclease(RNase A)with[V^(Ⅳ)O(8-HQ)_(2)]–a promising antitrypanosomal,antituberculosis,and antitumor vanadium complex(VC)formed by 8-hydroxyquinolinato(8-HQ)...Herein,we studied the interaction of bovine pancreatic ribonuclease(RNase A)with[V^(Ⅳ)O(8-HQ)_(2)]–a promising antitrypanosomal,antituberculosis,and antitumor vanadium complex(VC)formed by 8-hydroxyquinolinato(8-HQ),reaching IC_(50) values lower than cisplatin in HCT116 and A2780 cancer lines(0.99 and 0.96μM vs.15.0 and 3.4μM)and the minimum inhibitory concentration(MIC)of 3.7μM against strains of Mycobacterium tuberculosis.The system was investigated by spectroscopic(UV-vis absorption spectroscopy,circular dichroism,and electron paramagnetic resonance),computational(docking and DFT calculations),and X-ray crystallographic multi-technique approach.Crystallographic results at pH 5.1 demonstrate that upon interaction with the protein,one of the 8-HQ ligands is replaced by a water molecule and by Glu111 side-chain,resulting in the inhibition of the RNase A activity.EPR data confirm the formation of this adduct,with V^(Ⅳ) coordinated by 8-HQ and by the couples(Glu-COO^(−);H_(2)O)at pH 5.1,(His-N;H_(2)O)at pH 7.4,and(His-N;OH^(−))or(Ser/Thr-O^(−),H_(2)O)at pH 8.4.Computational data unveil the reasons behind the ligand exchange,not observed for similar complexes like[V^(Ⅳ)O(picolinato)_(2)]and not predicted by thermodynamic considerations.Overall,our results show that the VC–protein binding is defined not only by the thermodynamic stability of VC but also by other factors,such as the structure and steric requirements of the metal ligands,with important implications in the drug delivery strategies.展开更多
Here we report the binding of the diruthenium complex[Ru_(2)Cl(D-p-CNPhF)(O_(2)CCH_(3))_(3)]_(n)(D-p-CNPhF-=N,N’-bis(4-cyanophenyl)formamidinate)to the model protein bovine pancreatic ribonuclease(RNase A),investigat...Here we report the binding of the diruthenium complex[Ru_(2)Cl(D-p-CNPhF)(O_(2)CCH_(3))_(3)]_(n)(D-p-CNPhF-=N,N’-bis(4-cyanophenyl)formamidinate)to the model protein bovine pancreatic ribonuclease(RNase A),investigated for the first time by means of X-ray crystallography and Quantum Chemistry.The crystal structure reveals that the compound binds a histidine side chain with the diruthenium core anchored to RNase A at the axial site.展开更多
Herein,we present a detailed metalation process of the bovine pancreatic ribonuclease(RNase A)by a novel multitarget anti-cancer agent arsenoplatin-1,AP1,([Pt(μ-NHC(CH_(3))O)_(2)ClAs(OH)_(2)]),on the basis of quantum...Herein,we present a detailed metalation process of the bovine pancreatic ribonuclease(RNase A)by a novel multitarget anti-cancer agent arsenoplatin-1,AP1,([Pt(μ-NHC(CH_(3))O)_(2)ClAs(OH)_(2)]),on the basis of quantum chemical investigation,employing the B3LYP and M062x functionals and a large model of the active site.The proposed mechanism is consistent with the structural data.The role of water molecules in the active site is also analyzed.These studies revealed that the Nδof His119 binds platinum(Ⅱ),preserving the Pt-As bond.To better rationalize the different behavior of AP1(bound to His119)and the reference cisplatin(bound to Met29)towards the same target RNAse A,also these processes have been considered.The final platinated complex structure agrees well with the crystallographic one.Our results evidence that the metalation process takes place more favorably in water than in the protein environment in agreement with the nature of the protein binding pocket residues.展开更多
Ribonuclease E(RNase E)is central to bacterial RNA metabolism.In cyanobacteria,its activity is inhibited by RebA,a key mechanism for controlling cell morphology.Here,we demonstrate that rebA is essential for diazotrop...Ribonuclease E(RNase E)is central to bacterial RNA metabolism.In cyanobacteria,its activity is inhibited by RebA,a key mechanism for controlling cell morphology.Here,we demonstrate that rebA is essential for diazotrophic growth of Anabaena PCC 7120,a filamentous cyanobacterium capable of forming heterocysts-specialized nitrogen-fixing cells-upon nitrogen starvation.The rebA mutant strain(ΔrebA)showed severe growth defects in nitrogen-deprived conditions,despite forming more heterocysts than the wild type.With a GFP fusion strain,we show that RebA is transiently upregulated during heterocyst differentiation.Microscopic and ultrastructural analyses revealed thatΔrebA heterocysts accumulated abnormally large cyanophycin granules,while vegetative cells showed reduced pigment levels and disorganized thylakoid membranes,phenotypes indicative of a severe nitrogen deficiency response.However,esculin tracer diffusion and SepJ-GFP localization inΔrebA were comparable to the wild type,suggesting that cell-cell communication via septal junctions remains functional.Thus,the growth defect likely results from impaired degradation or mobilization of fixed nitrogen.Notably,theΔrebA phenotype could be rescued only by wild-type RebA,but not by variants unable to bind RNase E,indicating that RebA's function depends on its modulation of RNase E activity.Together,these findings reveal a key posttranscriptional mechanism linking RNase E regulation to heterocyst development and intercellular nutrient transfer,highlighting the importance of regulated RNA metabolism for diazotrophic growth.展开更多
Sequence-specific ribonuclease(RNase)P ribozymes can be engineered in vitro and are promising gene-targeting agents to knock down gene expression.In this study,we applied an RNase P ribozyme variant to hydrolyze the m...Sequence-specific ribonuclease(RNase)P ribozymes can be engineered in vitro and are promising gene-targeting agents to knock down gene expression.In this study,we applied an RNase P ribozyme variant to hydrolyze the mRNA of human cytomegalovirus(HCMV)major capsid protein(MCP),which is necessary for viral capsid formation and growth.Functional variant R668-F was about 100 times more efficient in slicing the MCP mRNA in vitro than M1-F,the ribozyme with a natural RNase P ribozyme sequence.In R668-F-expressing cells,a decrease of about 98%-99%in the expression of MCP was detected,and the virus production was reduced by 70,000 folds.However,the expression of inactive control ribozymes in cells resulted in a less than 10%decrease in MCP expression and no apparent decrease in HCMV growth.In cells observed with the ribozyme-mediated reduction of MCP expression,HCMV capsid formation and virus growth were inhibited and the expressions of other viral genes were unaffected.These findings provide the first direct evidence that ribozyme R668-F specifically inhibits MCP expression and blocks HCMV growth.Our results further suggest that the engineered RNase P ribozymes,including R668-F,may act as a novel general gene-targeting strategy to treat infections of viruses,including HCMV.展开更多
The islets of Langerhans orchestrate body responses to nutritional,neural,and hormonal stimuli,thus intricately maintaining blood glucose homeostasis and shaping the metabolic landscape[1].This specialized micro-organ...The islets of Langerhans orchestrate body responses to nutritional,neural,and hormonal stimuli,thus intricately maintaining blood glucose homeostasis and shaping the metabolic landscape[1].This specialized micro-organ is primarily composed ofα,β,δ,and PP cells,with the insulin-producingβcells being the dominant cell type[2].Abnormalities inβcell proliferation,mass,and function have been observed in various metabolic disorders[2,3].Thus,maintaining an optimalβcell proliferation capacity is essential not only for islet development and metabolic regulation but also for functional recovery and regeneration of the dysfunctionalβcells and islets[2].展开更多
RNA turnover plays critical roles in the regulation of gene expression and allows cells to respond rapidly to environmental changes.In bacteria,the mechanisms of RNA turnover have been extensively studied in the model...RNA turnover plays critical roles in the regulation of gene expression and allows cells to respond rapidly to environmental changes.In bacteria,the mechanisms of RNA turnover have been extensively studied in the models Escherichia coli and Bacillus subtilis,but not much is known in other bacteria.Cyanobacteria are a diverse group of photosynthetic organisms that have great potential for the sustainable production of valuable products using CO_(2)and solar energy.A better understanding of the regulation of RNA decay is important for both basic and applied studies of cyanobacteria.Genomic analysis shows that cyanobacteria have more than 10 ribonucleases and related proteins in common with E.coli and B.subtilis,and only a limited number of them have been experimentally investigated.In this review,we summarize the current knowledge about these RNAturnover-related proteins in cyanobacteria.Although many of them are biochemically similar to their counterparts in E.coli and B.subtilis,they appear to have distinct cellular functions,suggesting a different mechanism of RNA turnover regulation in cyanobacteria.The identification of new players involved in the regulation of RNA turnover and the elucidation of their biological functions are among the future challenges in this field.展开更多
基金financed by the National Science Centre, Poland (No. 2019/35/B/NZ2/02658 to P.J.).
文摘Ribonucleases (RNases), essential for RNA metabolism, are implicated in human diseases, including neurodevelopmental, developmental, hematopoietic and other dysfunctions through mutations that disrupt their enzymatic functions. Exploring RNase mutations across organisms offers insights into Mendelian diseases, facilitating molecular dissection of pathological pathways and therapeutic development. By employing model organisms, our analysis underscores the evolutionary conservation of RNase genes, facilitating deeper insights into disease mechanisms. These models are vital for uncovering rare molecular dysfunctions and potential therapeutic targets, demonstrating the effectiveness of integrated research approaches in addressing complex genetic disorders. Drawing from phylogenetic analyses, literature survey, and databases documenting the effects of human disease-causing mutations, the review highlights the significance and advantages of employing model organisms to study specific Mendelian disorders.
文摘Self-incompatibility is an intraspecific reproductive barrier to prevent self-fertilization inthe flowering plants. In many species, self-incompatibility is controlled by a single S locus with multiple alleles. So far, the only gene known in the S locus of the Solanaceae, Scrophulariaceae and Rosaceae encodes a class of ribonucleases, called self-incompatibility ribonucleases (S RNases), which have been shown to mediate stylar expression of self-incompatible reaction. As the first step to investigate their three-dimensional structure, we successfully expressed three biologically active S RNases of Antirrihnum (S2, S4 and S5) in Escherichia coli (E. coli). Their functional expressions caused no detrimental effect on host bacteria growth and provided a basis for a large scale preparation of S RNase proteins. Possible reasons for non-lethality of S RNases on E. coliare discussed.
基金The authors would like to thank Mr Shou-Xin Zhang and other members of the Research Center,Yuhuangding Hospital(Yantai,China)for technical assistance.
文摘To explore the functions of human ribonuclease 9(RNase 9),we constructed a mammalian fusion expression vector pcDNA-hRNase9,prepared recombinant human RNase 9-His fusion protein from HEK293T cells and determined its N-terminal amino acid sequences.According to the determined mature protein,recombinant human RNase 9 was prepared in E.coli.Ribonucleolytic activity and antibacterial activity of recombinant human RNase 9 were detected,and the distribution of human RNase 9 on tissues and ejaculated spermatozoa and in vitro capacitated spermatozoa were analyzed via indirect immunofluorescence assay.The results showed that recombinant human RNase 9 did not exhibit detectable ribonucleolytic activity against yeast tRNA,but exhibited antibacterial activity,in a concentration/time dependent manner,against E.coli.Immunofluorescent analyses showed that the predicted human RNase 9 was present throughout the epididymis,but not present in other tissues examined,and human RNase 9 was also present on the entire head and neck regions of human ejaculated spermatozoa and in vitro capacitated spermatozoa.These results suggest that human RNase 9 may play roles in host defense of male reproductive tract.
基金supported by the Central Public Interest Scientific Institution Basal Research Fund for the Chinese Academy of Agricultural Sciences(Grant No.:Y2021PT05)National Institute of Environmental Health Science Superfund Research Program(Grant No.:P42 ES004699)+1 种基金National Academy of Sciences(Subaward No.:2000009144)Ningbo Innovation Project for Agro-Products Quality and Safety(Grant No.:2019CXGC007).
文摘Balancing the risks and benefits of organophosphate pesticides(OPs)on human and environmental health relies partly on their accurate measurement.A highly sensitive fluorescence anti-quenching multi-residue bio-barcode immunoassay was developed to detect OPs(triazophos,parathion,and chlorpyrifos)in apples,turnips,cabbages,and rice.Gold nanoparticles were functionalized with monoclonal antibodies against the tested OPs.DNA oligonucleotides were complementarily hybridized with an RNA fluorescent label for signal amplification.The detection signals were generated by DNA-RNA hybridization and ribonuclease H dissociation of the fluorophore.The resulting fluorescence signal enables multiplexed quantification of triazophos,parathion,and chlorpyrifos residues over the concentration range of 0.01-25,0.01-50,and 0.1-50 ng/mL with limits of detection of 0.014,0.011,and 0.126 ng/mL,respectively.The mean recovery ranged between 80.3% and 110.8% with relative standard deviations of 7.3%-17.6%,which correlate well with results obtained by liquid chromatography-tandem mass spectrometry(LC-MS/MS).The proposed bio-barcode immunoassay is stable,reproducible and reliable,and is able to detect low residual levels of multi-residue OPs in agricultural products.
基金The work supported by the grant Ministry of Education of the Czech Republic with co-financing from the European Union(grant“KOROLID”,CZ.02.1.01/0.0/0.0/15_003/0000336)the Czech Academy of Sciences(RVO:60077344).
文摘Members of the Pathogenesis Related(PR)10 protein family have been identified in a variety of plant species and a wide range of functions ranging from defense to growth and development has been attributed to them.PR10 protein possesses ribonuclease(RNase)activity,interacts with phytohormones,involved in hormone-mediated signalling,afforded protection against various phytopathogenic fungi,bacteria,and viruses particularly in response to biotic and abiotic stresses.The resistance mechanism of PR10 protein may include activation of defense signalling pathways through possible interacting proteins involved in mediating responses to pathogens,degradation of RNA of the invading pathogens.Moreover,several morphological changes have been shown to accompany the enhanced abiotic stress tolerance.In this review,the possible mechanism of action of PR10 protein against biotic and abiotic stress has been discussed.Furthermore,our findings also confirmed that the in vivo Nitric oxide(NO)is essential for most of environmental abiotic stresses and disease resistance against pathogen infection.The proper level of NO may be necessary and beneficial,not only in plant response to the environmental abiotic stress,but also to biotic stress.The updated information on this interesting group of proteins will be useful in future research to develop multiple stress tolerance in plants.
基金supported by the program of key medical department of Jiangsu Province(No.XK17 200904)
文摘Objective：The association between ribonuclease L（RNASEL）gene polymorphisms and prostate cancer risk has been widely reported,but the results of these studies remained controversial and underpowered.We performed a meta-analysis of 28 studies to evaluate the association between Arg462Gln and Asp541Glu polymorphisms in the RNASEL gene and prostate cancer risk.Methods：Odds ratios（ORs）with 95%confidence intervals（CIs） were estimated to assess the association between RNASEL polymorphisms and prostate cancer risk.Results：A significantly increased prostate cancer risk was found for the Arg462Gln polymorphism in Africans（Gln/Gln vs Arg/Arg：OR=2.50,95%CI=1.28-4.87;Gln/Gln vs Gln/Arg＋Arg/Arg：OR=2.54,95%CI=1.30-4.95）,but not in Europeans and Asians.Additionally,the Asp541Glu polymorphism was associated with increased total prostate cancer risk（Glu-allele vs Asp-allele：OR=1.04,95%CI=1.01-1.07;Glu/Glu vs Asp/Asp：OR=1.22,95%CI= 1.03-1.46;Glu/Glu vs Glu/Asp＋Asp/Asp：OR=1.09,95%CI=1.02-1.16）.In the stratified analysis for the Asp541Glu polymorphism,there was a significantly increased prostate cancer risk in Africans and Europeans,and in hospital-based prostate cancer cases.Conclusion：The meta-analysis results showed evidence that RNASEL Arg462Gln and Asp541Glu polymorphisms are associated with prostate cancer risk and could be low-penetrance prostate cancer susceptibility biomarkers.
基金supported by grants from the National Key Research and Development Program of China(2022YFC2702604)the National Natural Science Foundation of China(82171608,82201773,and 81971447)+1 种基金the China Postdoctoral Science Foundation(2022M711119)the Scientific Research Foundation of the Health Committee of Hunan Province(B202301039323 and B202301039518).
文摘Male infertility is a major reproductive disorder,which is clinically characterized by highly heterogeneous phenotypes of abnormal sperm count or quality.To date,five male patients with biallelic loss-of-function(LOF)variants of PARN-like ribonuclease domain-containing exonuclease 1(PNLDC1)have been reported to experience infertility with nonobstructive azoospermia.The aim of this study was to identify the genetic cause of male infertility with oligo-astheno-teratozoospermia(OAT)in a patient from a Chinese Han family.Whole-exome and Sanger sequencing analyses identified a homozygous LOF variant(NM_173516.2,c.l42C>T,p.Gln48Ter)in PNLDC1.Hematoxylin and eosin staining revealed that the spermatozoa of the patient with OAT had an irregular head phenotype,including microcephaly,head tapering,and globozoospermia.Consistently,peanut agglutinin staining of the spermatozoa revealed a complete or partial loss of the acrosome.Furthermore,the disomy rate of chromosomes in the patient’s spermatozoa was significantly increased compared with that of a fertile control sample.We reported an LOF variant of the PNLDC1 gene responsible for OAT.
文摘BACKGROUND Bioinformatics analysis showed that the expression of the poly(A)-specific ribonuclease(PARN)gene in gastric cancer,head and neck squamous cell carcinoma,melanoma,cervical cancer and lung squamous cell carcinoma tissues was significantly higher than that in normal tissues and was associated with high stage and poor prognosis.The expression of the PARN gene in esophageal cancer(EC)tissue is also significantly higher than that in normal tissues,but the effect of PARN on the proliferation,migration and invasion of EC cells remains unclear.AIM To investigate the relationship between PARN and the proliferation,migration and invasion of EC cells.METHODS The EC tissues of 91 patients after EC surgery and 63 paired precancerous healthy tissues were collected.PARN mRNA levels were measured using a tissue microarray,and the PARN expression level was evaluated using immunohistochemistry to analyze the relationship between PARN expression and clinicopathologic features as well as the survival and prognosis of patients.In addition,the effects of PARN gene knockout on tumor cell proliferation,invasion and migration were studied by using shRNA during the in vitro culture of EC cell lines Eca-109 and TE-1,and the effects of the PARN gene on tumor growth in vivo were verified by a xenotransplantation nude mice model.RESULTS The expression of PARN in EC tissues was higher than that in adjacent normal tissues,and the level of PARN expression was significantly positively correlated with lymphatic metastasis.Patients with high PARN levels had poor overall survival.BIM,IGFBP-5 and p21 levels were significantly increased in the PARN knockout group,while the expression levels of the antiapoptotic proteins Survivin and sTNF-R1 were significantly decreased in the apoptotic antibody array data.In addition,the expression levels of Akt,p-Akt,PIK3CA and CCND1 in the downstream signaling pathway regulating EC progression were significantly decreased.The culture of EC cell lines confirmed that the apoptosis rate of EC cells was significantly increased,the growth and proliferation of tumor cells were significantly inhibited,and the invasion and migration ability of tumor cells were significantly decreased after PARN gene knockout.In vivo experiments of BALB/c nude mice transfected with Eca-109 cells expressing control shRNA(sh-NC)and PARN shRNA(sh-PARN)showed that the tumor volume and weight of nude mice treated with sh-PARN were significantly decreased compared with those of nude mice treated with sh-NC,indicating that PARN knockdown significantly inhibited tumor growth in vivo.CONCLUSION PARN has antiapoptotic effects on EC cells and promotes their proliferation,invasion and migration,which is associated with the development of EC and poor patient prognosis.PARN may become a potential target for the diagnosis,prognosis prediction and treatment of EC.
文摘Early stage expression of PR10 combined with phytoalexins contributed to Verticillium wilt resistance in cotton. In order to analysis the activities of PR10 proteins during pathogens’ infection, we cloned a Verticillium-induced PR10 (GbPR10-1) gene from cotton (Gossypium barbadense) and compared its expression patterns and domains with other PR10 proteins. Bioinformatics indicated that GbPR10-1 showed the lowest similarity with other 12 different PR10 genes in cotton (Upland and sea-island cotton). Expression profiles showed that GbPR10-1 gene instantly up-regulated after infection by V. dahliae in the sea-island cotton plants. GbPR10-1 was also induced by environmental stimulus including heat, submergence and salt, and ethylene but not by ABA and salicylic acid. The GbPR10-1 protein expressed in E. coli BL21 demonstrated that it had a low ribonuclease-like activity in vitro, and could inhibit V. dahliae hyphae growth but not its spores. Comparison analysis of GbPR10-1 (from resistant species) and GhPR10-1 (from susceptible species) responding to V. dahliae infection, only GbPR10-1 gene was strongly induced in the sea-island cotton plants (incompatible response), indicating that PR10-1 genes was linked to resistance signal. In summary, the earlier activation of GbPR10-1 gene, as the index of resistance response, would be aid to block
文摘Background: Viruses can cause different diseases in plants. To prevent viral infections, plants are treated with chemical compounds and antiviral agents. Chemical antiviral agents usually have narrow specificity, which limits their wide application. Alternative antiviral strategy is associated with the use of microbial enzymes, which are less toxic and are readily decomposed without accumulation of harmful substances. The aim of this work is to study the effect of Bacillus pumilus ribonuclease on various phytopathogenic viruses with specific focus on the ability of enzyme to eliminate them from plant explants in vitro. Materials and methods: Extracellular ribonuclease of B. pumilus is tested as an antiviral agent. To study the antiviral effect of RNase, depending on concentration and the time of application several plant-virus model systems are used. Virus detection is conducted by serological testing and RT-PCR. Results: Bacillus pumilus ribonuclease possesses antiviral activity against plant Rna-viruses RCMV (red clover mottle virus), PVX (Potato Virus X) and AMV (Alfalfa Mosaic Virus). The maximum inhibitory effect against actively replicating viruses is observed when plants are treated with the enzyme in the concentration of 100 ug/ml prior to infection. In case of local necrosis ribonuclease in the concentration of 1 ug/ml completely inhibits the development of RCMV virus on bean plants. The enzyme is able to penetrate plants and inhibit the development of viral infection, inhibiting effect for untreated surfaces decreased on average for 20%. It is also found that B. pumilus ribonuclease protects apical explants of sprouts of potato tubers from PVM and PVS viruses. Conclusion: B. pumilus ribonuclease possesses antiviral activity against plant Rna-viruses and produces viruses-free plants in the apical meristem culture.
文摘2-Chlorocyclohexa-2,5-diene-1,4-dione (CBQ) or 2-chloro1,4-benzquinone is one of the common metabolites of polycyclic aromatic hydrocarbons generated through industrial processes. This report describes the biological effects of CBQ toward ribonuclease A (RNase). We also investigated the inhibition of RNase modifications and the reactivity of CBQ toward selected amino acids. The study was carried out by incubating RNase or amino acids with CBQ in a concentration- and a time-dependent manner at 37°C and pH 7.0. SDS-PAGE results showed oligomerization as well as polymeric aggregation of RNase when incubated with CBQ as early as in 10 min. CBQ-induced RNase modifications were inhibited in the presence of NADH or ascorbic acid. CBQ reactivity toward selected amino acids was also evaluated by determining the second-order rate constants for the reactions of CBQ with selected amino acids. It was found that the reactivity toward CBQ decreased in the order of lysine > threonine > serine >> aspartate > cysteine.
基金supported by Fondazione di Sardegna(grant FdSGarribba2017)Spanish MINECO’Juan de la Cierva program,FJC2019-039135-I.
文摘Herein,we studied the interaction of bovine pancreatic ribonuclease(RNase A)with[V^(Ⅳ)O(8-HQ)_(2)]–a promising antitrypanosomal,antituberculosis,and antitumor vanadium complex(VC)formed by 8-hydroxyquinolinato(8-HQ),reaching IC_(50) values lower than cisplatin in HCT116 and A2780 cancer lines(0.99 and 0.96μM vs.15.0 and 3.4μM)and the minimum inhibitory concentration(MIC)of 3.7μM against strains of Mycobacterium tuberculosis.The system was investigated by spectroscopic(UV-vis absorption spectroscopy,circular dichroism,and electron paramagnetic resonance),computational(docking and DFT calculations),and X-ray crystallographic multi-technique approach.Crystallographic results at pH 5.1 demonstrate that upon interaction with the protein,one of the 8-HQ ligands is replaced by a water molecule and by Glu111 side-chain,resulting in the inhibition of the RNase A activity.EPR data confirm the formation of this adduct,with V^(Ⅳ) coordinated by 8-HQ and by the couples(Glu-COO^(−);H_(2)O)at pH 5.1,(His-N;H_(2)O)at pH 7.4,and(His-N;OH^(−))or(Ser/Thr-O^(−),H_(2)O)at pH 8.4.Computational data unveil the reasons behind the ligand exchange,not observed for similar complexes like[V^(Ⅳ)O(picolinato)_(2)]and not predicted by thermodynamic considerations.Overall,our results show that the VC–protein binding is defined not only by the thermodynamic stability of VC but also by other factors,such as the structure and steric requirements of the metal ligands,with important implications in the drug delivery strategies.
基金supported by the Human Frontier Science Program(Grant Number RGY0056/2022)A.M.thanks MIUR PRIN 2022-Cod.2022JMFC3X,“Protein Metalation by Anticancer Metal-based Drugs”for financial support.Comunidad de Madrid(Project S2017/BMD-3770-CM)+2 种基金Complutense University of Madrid(GRFN32/23 and Program PR3/23)are gratefully acknowledged for financial supportA.T.also acknowledges the Complutense University for a Predoctoral Grant(CT63/19-CT64/19)Research Stay Grant(EB25/22)and the Spanish Ministry of Science and Innovation for a Postgraduate Fellowship at Residencia de Estudiantes(2021-2022)。
文摘Here we report the binding of the diruthenium complex[Ru_(2)Cl(D-p-CNPhF)(O_(2)CCH_(3))_(3)]_(n)(D-p-CNPhF-=N,N’-bis(4-cyanophenyl)formamidinate)to the model protein bovine pancreatic ribonuclease(RNase A),investigated for the first time by means of X-ray crystallography and Quantum Chemistry.The crystal structure reveals that the compound binds a histidine side chain with the diruthenium core anchored to RNase A at the axial site.
文摘Herein,we present a detailed metalation process of the bovine pancreatic ribonuclease(RNase A)by a novel multitarget anti-cancer agent arsenoplatin-1,AP1,([Pt(μ-NHC(CH_(3))O)_(2)ClAs(OH)_(2)]),on the basis of quantum chemical investigation,employing the B3LYP and M062x functionals and a large model of the active site.The proposed mechanism is consistent with the structural data.The role of water molecules in the active site is also analyzed.These studies revealed that the Nδof His119 binds platinum(Ⅱ),preserving the Pt-As bond.To better rationalize the different behavior of AP1(bound to His119)and the reference cisplatin(bound to Met29)towards the same target RNAse A,also these processes have been considered.The final platinated complex structure agrees well with the crystallographic one.Our results evidence that the metalation process takes place more favorably in water than in the protein environment in agreement with the nature of the protein binding pocket residues.
基金supported by the National Natural Science Foundation of China(32070037)the Knowledge Innovation Program of Wuhan-Basic Research(2023020201010087)+1 种基金the China Postdoctoral Science Foundation(2025M772687)the Postdoctoral Fellowship Program of CPSF(GZC20241873)。
文摘Ribonuclease E(RNase E)is central to bacterial RNA metabolism.In cyanobacteria,its activity is inhibited by RebA,a key mechanism for controlling cell morphology.Here,we demonstrate that rebA is essential for diazotrophic growth of Anabaena PCC 7120,a filamentous cyanobacterium capable of forming heterocysts-specialized nitrogen-fixing cells-upon nitrogen starvation.The rebA mutant strain(ΔrebA)showed severe growth defects in nitrogen-deprived conditions,despite forming more heterocysts than the wild type.With a GFP fusion strain,we show that RebA is transiently upregulated during heterocyst differentiation.Microscopic and ultrastructural analyses revealed thatΔrebA heterocysts accumulated abnormally large cyanophycin granules,while vegetative cells showed reduced pigment levels and disorganized thylakoid membranes,phenotypes indicative of a severe nitrogen deficiency response.However,esculin tracer diffusion and SepJ-GFP localization inΔrebA were comparable to the wild type,suggesting that cell-cell communication via septal junctions remains functional.Thus,the growth defect likely results from impaired degradation or mobilization of fixed nitrogen.Notably,theΔrebA phenotype could be rescued only by wild-type RebA,but not by variants unable to bind RNase E,indicating that RebA's function depends on its modulation of RNase E activity.Together,these findings reveal a key posttranscriptional mechanism linking RNase E regulation to heterocyst development and intercellular nutrient transfer,highlighting the importance of regulated RNA metabolism for diazotrophic growth.
基金supported by a Block Grant from the Graduate Division of the University of California at Berkeley.
文摘Sequence-specific ribonuclease(RNase)P ribozymes can be engineered in vitro and are promising gene-targeting agents to knock down gene expression.In this study,we applied an RNase P ribozyme variant to hydrolyze the mRNA of human cytomegalovirus(HCMV)major capsid protein(MCP),which is necessary for viral capsid formation and growth.Functional variant R668-F was about 100 times more efficient in slicing the MCP mRNA in vitro than M1-F,the ribozyme with a natural RNase P ribozyme sequence.In R668-F-expressing cells,a decrease of about 98%-99%in the expression of MCP was detected,and the virus production was reduced by 70,000 folds.However,the expression of inactive control ribozymes in cells resulted in a less than 10%decrease in MCP expression and no apparent decrease in HCMV growth.In cells observed with the ribozyme-mediated reduction of MCP expression,HCMV capsid formation and virus growth were inhibited and the expressions of other viral genes were unaffected.These findings provide the first direct evidence that ribozyme R668-F specifically inhibits MCP expression and blocks HCMV growth.Our results further suggest that the engineered RNase P ribozymes,including R668-F,may act as a novel general gene-targeting strategy to treat infections of viruses,including HCMV.
基金supported by the following grants:the National Natural Science Foundation of China(32101015 and 82270670)Fundamental Research Funds for the Central Universities(2021QNA7009)+1 种基金Leading Innovative and Entrepreneur Team Introduction Program of Zhejiang(2021R01012)Leading Innovation and Entrepreneur Team of Hangzhou(TD2020006).
文摘The islets of Langerhans orchestrate body responses to nutritional,neural,and hormonal stimuli,thus intricately maintaining blood glucose homeostasis and shaping the metabolic landscape[1].This specialized micro-organ is primarily composed ofα,β,δ,and PP cells,with the insulin-producingβcells being the dominant cell type[2].Abnormalities inβcell proliferation,mass,and function have been observed in various metabolic disorders[2,3].Thus,maintaining an optimalβcell proliferation capacity is essential not only for islet development and metabolic regulation but also for functional recovery and regeneration of the dysfunctionalβcells and islets[2].
基金supported by the National Natural Science Foundation of China(grant No.32070037)the Featured Institute Service Project from the Institute of Hydrobiology,the Chinese Academy of Sciences(grant No.Y85Z061601)the German Science Foundation(DFG)research training group BioInMe 322977937/GRK2344 and grant HE 2544/14-2(to Wolfgang R.Hess).
文摘RNA turnover plays critical roles in the regulation of gene expression and allows cells to respond rapidly to environmental changes.In bacteria,the mechanisms of RNA turnover have been extensively studied in the models Escherichia coli and Bacillus subtilis,but not much is known in other bacteria.Cyanobacteria are a diverse group of photosynthetic organisms that have great potential for the sustainable production of valuable products using CO_(2)and solar energy.A better understanding of the regulation of RNA decay is important for both basic and applied studies of cyanobacteria.Genomic analysis shows that cyanobacteria have more than 10 ribonucleases and related proteins in common with E.coli and B.subtilis,and only a limited number of them have been experimentally investigated.In this review,we summarize the current knowledge about these RNAturnover-related proteins in cyanobacteria.Although many of them are biochemically similar to their counterparts in E.coli and B.subtilis,they appear to have distinct cellular functions,suggesting a different mechanism of RNA turnover regulation in cyanobacteria.The identification of new players involved in the regulation of RNA turnover and the elucidation of their biological functions are among the future challenges in this field.
